<i>Cladosporium herbarum</i> Extract Characterized by Means of Quantitative Immunoelectrophoretic Methods with Special Attention to Immediate Type Allergy

Abstract

Freeze-dried extract of Cladosporium herbarum Link ex Fr. was obtained by growing, harvesting, extracting, centrifuging, dialysing and freeze-drying. Quantitative immunoelectrophoresis using rabbit antibodies revealed the extraction procedure to be reproducible and the extract to be composed of 57 antigens, none of which originated from the substrate used in the growth. The molecular weight distribution and the approximate molecular weight of some antigens of C. herbarum were obtained using gel filtration. The pI distribution and the approximate pIs of a few distinct antigens of C. herbarum were obtained by isoelectric focusing. Preliminary identification of 4 allergens from C. herbarum was performed by means of CRIE (crossed radioimmunoelectrophoresis).

Partial Purification and Characterization of Two <i>Cladosporium herbarum </i>Allergens

“…The present work verified a pre vious report that most antigenic proteins in C. herbarum have molecular weights even higher than 70,000 daltons [6].…”

“…Lpwenstein et al [6] used quantitative immunoelectrophoretic techniques to char acterize an extract of C. herbarum and showed the distribution of molecular weights and isoelectric points of the anti gens. Preliminary attempts to identify the allergens in this extract by means of crossed radioimmunoelectrophoresis (CR1E) were not conclusive, due to the low number of patient sera employed.…”

Crossed Radioimmunoelectrophoretic Studies of Distinct Allergens in Two Extracts of <i>Cladosporium herbaru</i><i>m</i>

“…A CIE reference pattern of C. herbarum showing 57 precipitates has been published previously [15]. In the CIE reference pat terns of Ch-Allergon and Ch-77.…”

“…Aller gen extracts of molds have also been report ed to contain irritants restricting their clini cal value [7]. An extract of C. herbarum (Ch-76) has been characterized previously using quantitative immunoelectrophoretic techniques, but preliminary attempts to identify the allergens in this extract were not conclusive due to the low number of patient sera employed [15]. Only moderate quanti tative differences were then observed when two different batches of the mold were com pared.…”

“…When the Ch-Allergon and Ch-77 extracts and antisera were cross-combined in CIE, the number of detectable precipitates was drastically reduced. Since the Ch-Allergon and Ch-77 extracts were prepared identical ly according to a procedure which has been shown to give reproducible results [15], this suggested that different batches from one manufacturer vary considerably less than extracts from different manufacturers. AlIergon AB and Allergologisk Laboratorium A/'S employ different strains of C. herbarum and different growth procedures.…”